Single core genome sequencing for detection of both Borrelia burgdorferi Sensu Lato and relapsing fever Borrelia species

dc.contributor.authorLee, Sin Hang
dc.contributor.authorHealy, John Eoin
dc.contributor.authorLambert, John S.
dc.date.accessioned2019-10-29T05:36:54Z
dc.date.available2019-10-29T05:36:54Z
dc.date.issued2019-05-20
dc.description.abstractLyme disease, initially described as Lyme arthritis, was reported before nucleic-acid based detection technologies were available. The most widely used diagnostic tests for Lyme disease are based on the serologic detection of antibodies produced against antigens derived from a single strain of Borrelia burgdorferi. The poor diagnostic accuracy of serological tests early in the infection process has been noted most recently in the 2018 Report to Congress issued by the U.S. Department of Health and Human Services Tick-Borne Disease Working Group. Clinical Lyme disease may be caused by a diversity of borreliae, including those classified as relapsing fever species, in the United States and in Europe. It is widely accepted that antibiotic treatment of Lyme disease is most successful during this critical early stage of infection. While genomic sequencing is recognized as an irrefutable direct detection method for laboratory diagnosis of Lyme borreliosis, development of a molecular diagnostic tool for all clinical forms of borreliosis is challenging because a “core genome” shared by all pathogenic borreliae has not yet been identified. After a diligent search of the GenBank database, we identified two highly conserved segments of DNA sequence among the borrelial 16S rRNA genes. We further developed a pair of Borrelia genus-specific PCR primers for amplification of a segment of borrelial 16S rRNA gene as a “core genome” to be used as the template for routine Sanger sequencing-based metagenomic direct detection test. This study presented examples of base-calling DNA sequencing electropherograms routinely generated in a clinical diagnostic laboratory on DNA extracts of human blood specimens and ticks collected from human skin bites and from the environment. Since some of the tick samples tested were collected in Ireland, borrelial species or strains not known to exist in the United States were also detected by analysis of this 16S rRNA “core genome”. We recommend that hospital laboratories located in Lyme disease endemic areas begin to use a “core genome” sequencing test to routinely diagnose spirochetemia caused by various species of borreliae for timely management of patients at the early stage of infection.en
dc.description.statusPeer revieweden
dc.description.versionPublished Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleid1779en
dc.identifier.citationLee, S.H., Healy, J.E. and Lambert, J.S., 2019. Single Core Genome Sequencing for Detection of both Borrelia burgdorferi Sensu Lato and Relapsing Fever Borrelia Species. International journal of environmental research and public health, 16(10), 1779 (22 pp.) DOI:10.3390/ijerph16101779en
dc.identifier.doi10.3390/ijerph16101779en
dc.identifier.eissn1660-4601
dc.identifier.endpage22en
dc.identifier.issn1661-7827
dc.identifier.issued10en
dc.identifier.journaltitleInternational journal of environmental research and public healthen
dc.identifier.startpage1en
dc.identifier.urihttps://hdl.handle.net/10468/8904
dc.identifier.volume16en
dc.language.isoenen
dc.publisherMDPIen
dc.relation.urihttps://www.mdpi.com/1660-4601/16/10/1779
dc.rights© 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) licenseen
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subjectLyme diseaseen
dc.subjectCore genomeen
dc.subjectDNA sequencingen
dc.subjectBorreliosisen
dc.subjectRelapsing fever borreliaeen
dc.subjectMetagenomic diagnosisen
dc.subjectBorreliaen
dc.subjectGenus-specific PCR primersen
dc.subjectSecond PCRen
dc.subjectDirect detectionen
dc.subjectSanger sequencingen
dc.titleSingle core genome sequencing for detection of both Borrelia burgdorferi Sensu Lato and relapsing fever Borrelia speciesen
dc.typeArticle (peer-reviewed)en
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